Reagent
(1) Coating buffer (PH9.60.05M carbonate buffer):
Na2CO3 1.59g
NaHCO3 2.93g
Add distilled water to 1000ml
(2) Washing buffer (PH7.4PBS): 0.15M
KH2PO4 0.2g
Na2HPO4 · 12H2O 2.9g
NaCl 8.0g
KCl 0.2g
Tween-20 0.05% 0.5ml
Add distilled water to 1000ml
(3) Diluent:
Bovine serum albumin (BSA) 0.1 g
Add wash buffer to 100ml
Or use sheep serum, rabbit serum and other serums to mix with 5-10% of the washing solution.
(4) Stop solution (3M H2SO4):
Distilled water 178.3ml, 21.7ml of concentrated sulfuric acid (98%) was added dropwise.
(5) Substrate buffer (PH5.0 phosphate date citric acid):
0.2MNa2HPO4 (28.4g / L) 25.7ml
0.1M citric acid (19.2g / L) 24.3ml
Add 50ml of distilled water.
(6) TMB (tetramethylbenzidine) use solution:
TMB (10mg / 5ml absolute ethanol) 0.5ml
Substrate buffer (PH5.5) 10ml
0.75% H2O2 32μl
(7) ABTS solution:
ABTS 0.5mg
Substrate buffer (PH5.5) 1ml
3% H2O2 2μl
(8) Antigen, antibody and enzyme labeled antibody.
(9) Normal human serum and positive control serum.
equipment
(1) 40- or 96-well polystyrene plastic plate (abbreviated as enzyme-labeled plate), ELISA detector, 50μl and 100μl sampler, plastic dripper, small towel, washing bottle.
(2) Small beakers, glass rods, test tubes, straws and measuring cylinders.
(3) 4 ℃ refrigerator, 37 ℃ incubator.
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